Team:Paris/Production testing

From 2009.igem.org

(Difference between revisions)
(→Part I : Testing delay phase I)
(→Part I : Testing delay phase I)
Line 175: Line 175:
RBS-YFP-ter  E/X
RBS-YFP-ter  E/X
-
|bgcolor="#BCCDFD"|'''Digestion:'''
+
|bgcolor="#BCCDFD"|-
|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
Line 201: Line 201:
|bgcolor="#E0E3FE"|'''Ligation:'''  
|bgcolor="#E0E3FE"|'''Ligation:'''  
|bgcolor="#CBD1FD"|'''Ligation:'''  
|bgcolor="#CBD1FD"|'''Ligation:'''  
-
|bgcolor="#BCCDFD"|-
+
|bgcolor="#BCCDFD"|'''Ligation:'''
|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
-
|bgcolor="#F8F8F8"|-
+
|bgcolor="#F8F8F8"|'''Colony PCR :'''
 +
ok
|bgcolor="#E0E3FE"|'''Colony PCR :'''
|bgcolor="#E0E3FE"|'''Colony PCR :'''
Line 211: Line 212:
|bgcolor="#CBD1FD"|'''Colony PCR :'''
|bgcolor="#CBD1FD"|'''Colony PCR :'''
-
-
+
ok
|bgcolor="#BCCDFD"|'''Colony PCR:'''
|bgcolor="#BCCDFD"|'''Colony PCR:'''
-
-
+
ok
|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
|bgcolor="#F8F8F8"|'''Miniprep:'''
|bgcolor="#F8F8F8"|'''Miniprep:'''
-
ok
+
clone
|bgcolor="#E0E3FE"|'''Miniprep:'''
|bgcolor="#E0E3FE"|'''Miniprep:'''
-
ok
+
clone
|bgcolor="#CBD1FD"|'''Miniprep:'''
|bgcolor="#CBD1FD"|'''Miniprep:'''
-
 
+
clone
-
ok
+
|bgcolor="#BCCDFD"|-
|bgcolor="#BCCDFD"|-
Line 230: Line 230:
|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
|bgcolor="#F8F8F8"|'''Sequencing :'''
|bgcolor="#F8F8F8"|'''Sequencing :'''
-
 
+
ok
-
-
+
|bgcolor="#E0E3FE"|'''Sequencing :'''
|bgcolor="#E0E3FE"|'''Sequencing :'''
-
 
+
ok
-
-
+
|bgcolor="#CBD1FD"|'''Sequencing :'''
|bgcolor="#CBD1FD"|'''Sequencing :'''

Revision as of 01:13, 22 October 2009

iGEM > Paris > WetLab > Production


Contents

WetLab - Production

  1. The delay
    1. Part I : Testing delay phase I
    2. Part I : Functional testing of the part I
    3. Part II : Testing delay phase II
    4. Part II : Functional testing of the part II
  2. The destabilization of the membrane

The delay

We separate the construction on two plasmids : Part I and Part II.

Part I : Testing delay phase I

The green color means the experiment was a success


plasmid = PSB3T5



Experiments ran :


Column 1 Column 2 Column 3 Column 4
PCR :

pBAD: plate igem


LacI: plate igem


Terminator: plate igem


RBS: plate igem


YFP: plate igem

- - -
Verification on gel :

ok

- - -
Purification on gel :

ok

- - -



Digestion:

pBAD S/P


lacI E/S


terminator E/X


RBS S/P


YFP X/P

Digestion:

pBAD-LacI E/S


RBS-YFP E/S

Digestion:

RBS-YFP-ter E/X

-
Verification digestion:

ok

Verification digestion:

ok

Verification digestion:

ok

-


Ligation:

pBAD-LacI

RBS-YFP

Ligation: Ligation: Ligation:
Colony PCR :

ok

Colony PCR :

ok

Colony PCR :

ok

Colony PCR:

ok

Miniprep:

clone

Miniprep:

clone

Miniprep:

clone

-
Sequencing :

ok

Sequencing :

ok

Sequencing :

-

Sequencing :

-

Stock glycerol:


Stock glycerol

Cly A(Cter)-(Nter)RFP: S72(clone 8)

Stock glycerol

-

Stock glycerol

-

Part I : Functional testing of the part I

PBAD LacI LVA- YFP was transforme into Top ten Bacteria and grew on medium :


1% arabinose

1% glucose

LB pure


Part II : Testing delay phase II

The green color means the experiment was a success


plasmid =



Experiments ran :


Column 1 Column 2 Column 3 Column 4
PCR :

mRFP Cterm matrix : pSB1A3 Oligo :O59 and O60 TM :


mRFP Nterm plasmid : pSB1A3 Oligo :O57 and O58 TM :

PCR : PCR : PCR :
Verification on gel :

ok

Verification on gel :

-

Verification on gel :

-

Verification on gel :

-

Purification on gel :

ok

Purification on gel :

-

Purification on gel :

-

Purification on gel :

-


Digestion:

TatABCE

Digestion: Digestion: Digestion:
Verification digestion:

ok

Verification digestion:

-

Verification digestion:

-

Verification digestion:

-


Ligation:

TatABCE

Ligation: Ligation: Ligation:
Colony PCR:

ok

Colony PCR :

-

Colony PCR:

-

Colony PCR :

-

Miniprep:

STOPPED

Miniprep:

-

Miniprep:

-

Miniprep:

-

Sequencing :

-

Sequencing :

-

Sequencing :

-

Sequencing :

-

Stock glycerol:

-

Stock glycerol

-

Stock glycerol

-

Stock glycerol

-

Part II : Functional testing of the part II

Ptet RFP Plac TetR was tranforme into Top10 bacteria and grew on media :


0,1% IPTG .

1% glucose.

LB pure.



The destabilization of the membrane

The green color means the experiment was a success


plasmid = PSB3T5



Experiments ran :


Column 1 Column 2 Column 3 Column 4
PCR :

TatABCE matrix :

PCR : PCR : PCR :
Verification on gel :

ok

Verification on gel :

-

Verification on gel :

-

Verification on gel :

-

Purification on gel :

ok

Purification on gel :

-

Purification on gel :

-

Purification on gel :

-


Digestion:

TatABCE

Digestion: Digestion: Digestion:
Verification digestion:

ok

Verification digestion:

-

Verification digestion:

-

Verification digestion:

-


Ligation:

TatABCE

Ligation: Ligation: Ligation:
Colony PCR :

ok

Colony PCR:

-

Colony PCR:

-

Colony PCR:

-

Miniprep:

STOPPED

Miniprep:

-

Miniprep:

-

Miniprep:

-

Sequencing :

-

Sequencing :

-

Sequencing :

-

Sequencing :

-

Stock glycerol:

-

Stock glycerol

-

Stock glycerol

-

Stock glycerol

-



Retrieved from "http://2009.igem.org/Team:Paris/Production_testing"